CHARACTERIZATION OF BETA RESISTALACTAM AND COLISTIN RESISTANT GRAM-NEGATIVE BACTERIA FROM ABATTOIR AND POULTRY EFFLUENTS WITHIN ABAKALIKI METROPOLIS, EBONYI STATE, NIGERIA.

SOURCE:

Faculty: Pharmaceutical Sciences
Department: Pharmaceutical Microbiology & Biotechnology

CONTRIBUTORS:

Nwakaeze, E. A.
Esimone, C. O.
Iroha, I. R.

ABSTRACT:

This study was designed to isolate and characterise antibiotics resistant genes fromEscherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae from abattoir and poultry effluents in Abakaliki metropolis in Ebonyi State Nigeria. With ethical approval, 354 effluent samples from both abattoir and poultry sites were collected and screened using microbiological techniques. Test organisms were characterized and identified phenotypically using biochemical test and Gram staining. Antimicrobial susceptibility testing was carried out using the Kirby - Bauer technique. Production of Extended-spectrum beta-lactamase (ESBL), Metallo beta-lactamase (MBL), AmpC and colistin resistance were determined phenotypically. Using specific primers, multiplex polymerase chain reaction was used to amplify resistance genes molecularly. All the effluent samples analysed had growth of Escherichia coli (46 %), Pseudomonas aeruginosa (29.0 %) and Klebsiella pneumoniae (25.0 %) from both abattoir and poultry effluents harbour resistance genes to antibiotics in the range 70 – 100 % from both sample source as shown in our study. Overall results from resistant enzyme production showed that 69.0 % of Escherichia coli isolated produces AmpC, 46.0 % produce ESBL, 62.0 % produce mcrenzymes and 46 produce MBL, Klebsiella speciesshowed 61.0 % for AmpC, 72.0 % for ESBL, 85.0 % for mcr and 31.0 % for MBL and Pseudomonas aeruginosa showed 57.0 % for AmpC, 64.0 % for ESBL, 72.0 % for mcrand 61.0 % for MBL. Multi-antibiotics resistance index (MARI) ranged from 0.4 – 1 in all the test isolates. Successful transconjugant strains showed resistance to CAZ, CTX, IPM, OB, FEP, CIP, FOX, CT, CRO, MEM and AMC. Molecular detection of the resistance genes revealed that TEM and CTX -M were more predominant among ESBL. IMP – 1 gene dominated among the MBL genes but only one isolate showed presence of IMP – 2 gene. FOX – 1, CMY – 1 and ACC were the predominantAmpC genes. Furthermore, mcr – 1 gene was detected in the isolates. This study is apparently the first to report IMP – 1, IMP – 2, ACC and mcr – 1 gene among isolates from abattoir and poultry in Ebonyi State and first to report ACC and IMP – 2 genes in South-Eastern Nigeria. However, unfortunately, resistance to this important safe and efficient classes of antibiotic is increasing worldwide. The resistance genetic elements were either encoded in the plasmid or chromosome and thus might be vertically propagated or acquired horizontally from the environment. Thus, appropriate hygiene and antibiotic usage should be intensified through public awareness and training of both livestock and poultry farmer. Also, stringent measures should be applied in access to antibiotic in non-clinical sectors to avoid propagation of resistant strains.